Polymorphisms of HIV-2 integrase and selection of resistance to raltegravir
Polymorphisms of HIV-2 integrase and selection of resistance to raltegravir
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DOI：
10.1186/1742-4690-7-98
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发表时间：
2010-11-29
期刊：
影响因子：
3.3
通讯作者：
Ruelle, Jean
中科院分区：
文献类型：
--
作者：
Bercoff, Danielle Perez;Triqueneaux, Perrine;Ruelle, Jean
背景：人类免疫缺陷病毒2型对某些抗逆转录病毒药物具有天然耐药性，限制了HIV-2感染患者的治疗选择。包括整合酶抑制剂（INI）在内的治疗方案似乎是有效的，但关于HIV-2整合酶（IN）多态性和耐药途径的数据很少。材料与方法：对45例hiv -2感染者的整合酶编码序列进行测序，并与ROD （A组）或EHO （B组）参考菌株比对，分析多态性或保守性位点。为了选择抗雷替格拉韦（raltegravir， RAL）的体外变异，ROD菌株在不断增加的次优RAL浓度下连续培养。所选变异的表型使用MTT测定法进行评估。结果：我们描述了45例HIV-2临床分离株的整合酶基因多态性。67%的整合酶残基是保守的。HHCC锌配位基序、催化三联体DDE基序和参与in - dna结合和正确定位的AA与HIV-1相比高度保守且不变，而n端结构域、二聚体界面和c端LEDGF结合结构域的连接残基与HIV-1相比高度保守但不同。在1例接受arv治疗的INI初发患者的病毒群体中检测到N155 H INI耐药相关突变（RAM），在36例和38例患者的样本中分别检测到72I和201I多态性。没有检测到其他已知的INI RAM。在体外RAL选择压力下，选择了一个携带Q91R+I175M突变的ROD变异体。Q91R和I175M突变同时出现，并赋予表型抗性（IC50增加13倍）。所有临床分离株均不存在Q91R+I175M组合。三维建模表明，残基91位于酶表面，位于含有DDE催化三元体的口袋的入口，并且添加正电荷（GIn到Arg）可能会损害IN-RAL的亲和力。结论：描述了45例初次感染i患者的HIV-2多态性。保守区域以及HIV-2 IN多态性的频率与HIV-1相当。两个新的突变（Q91R和I175M）在体外获得了对RAL的高耐药性，这可能会影响治疗结果。
Background: Human Immunodeficiency Virus type 2 is naturally resistant to some antiretroviral drugs, restricting therapeutic options for patients infected with HIV-2. Regimens including integrase inhibitors (INI) seem to be effective, but little data on HIV-2 integrase (IN) polymorphisms and resistance pathways are available.Materials and methods: The integrase coding sequence from 45 HIV-2-infected, INI-naive, patients was sequenced and aligned against the ROD (group A) or EHO (group B) reference strains and polymorphic or conserved positions were analyzed.To select for raltegravir (RAL)-resistant variants in vitro, the ROD strain was cultured under increasing sub-optimal RAL concentrations for successive rounds. The phenotype of the selected variants was assessed using an MTT assay.Results: We describe integrase gene polymorphisms in HIV-2 clinical isolates from 45 patients. Sixty-seven percent of the integrase residues were conserved. The HHCC Zinc coordination motif, the catalytic triad DDE motif, and AA involved in IN-DNA binding and correct positioning were highly conserved and unchanged with respect to HIV-1 whereas the connecting residues of the N-terminal domain, the dimer interface and C-terminal LEDGF binding domain were highly conserved but differed from HIV-1. The N155 H INI resistance-associated mutation (RAM) was detected in the virus population from one ARV-treated, INI-naive patient, and the 72I and 201I polymorphisms were detected in samples from 36 and 38 patients respectively. No other known INI RAM was detected.Under RAL selective pressure in vitro, a ROD variant carrying the Q91R+I175M mutations was selected. The Q91R and I175M mutations emerged simultaneously and conferred phenotypic resistance (13-fold increase in IC50). The Q91R+I175M combination was absent from all clinical isolates. Three-dimensional modeling indicated that residue 91 lies on the enzyme surface, at the entry of a pocket containing the DDE catalytic triad and that adding a positive charge (GIn to Arg) might compromise IN-RAL affinity.Conclusions: HIV-2 polymorphisms from 45 INI-naive patients are described. Conserved regions as well as frequencies of HIV-2 IN polymorphisms were comparable to HIV-1. Two new mutations (Q91R and I175M) that conferred high resistance to RAL were selected in vitro, which might affect therapeutic outcome.